LOCUS       sequence-175379-        7993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Halo-His6 tagged Nrf2 for E.coli expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7993)
  AUTHORS   Parvez S, Fu Y, Li J, Long MJ, Lin H, Lee DK, Hu GS, Aye Y
  TITLE     Substoichiometric Hydroxynonenylation of a Single Protein 
            Recapitulates Whole-Cell-Stimulated Antioxidant Response.
  JOURNAL   J Am Chem Soc. 2014 Dec 31.
  PUBMED    25544059
REFERENCE   2  (bases 1 to 7993)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Am Chem 
            Soc. 2014 Dec 31."
COMMENT     Sequence Label: pet28a-His6-Halo-TEV-Nrf2
FEATURES             Location/Qualifiers
     source          1..7993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(18..38)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(45..935)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             complement(939..956)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(966..983)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1002..1024
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1039..1063
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(1063..1082)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1064..1082)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1166..1185)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1391..1468
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1469..2551
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1488..1507)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3360..3551
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3545..3567)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3653..3792
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3708..3727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3807..3824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3978..4566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4058..4077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4677..4696)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4688..5503
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4688..5503
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4761..4780)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5595..6050)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5732..5753)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5944..5963
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(6087..6134)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     6130..6148
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 aagtccatca tgccagagcc ctggaaatac aagttttccc ctgagccgga aatctcgagc
       61 gtcgacagcc agcgcgcgat ctcgctgccg atcaggtccg ggttgtcttc ttgcagcaga
      121 ttcagacccg ggccgatgtc cacagccttg cagttaggca ggcttttggc caggcgagcg
      181 gcttcggccg gtgggatcag aacgcctggg gtgccccaga acagcagctt cgggacaggg
      241 gactggtgca gccagtccat gtattcttcg accagcgcga cgatgttcgc tggctcaccg
      301 gcgattggca gctcgtttgg gaagcgccac agtggctcgc ggtcaacagg attcaggaac
      361 ggctcgcggt aatggtccat ctcgacttca gtcagcgggc ggacgacacc catcggcagc
      421 gtaccctcga taaaaacgtt ctgatcgatg atcagcttgc ggccgacgtc ggtggtgcgg
      481 aaggcctgga aggtctcgcg ggcaaattct ggccattcgt cccaggtcgg gatagggcgg
      541 atgaactcca taaatgcaat acctttgacg cgctctggat tgcgcttggc ccagtggaaa
      601 cccagagcgg agccccagtc gtgaatgacc aggacgacct cttccagacc cagggcttcg
      661 atgaaggcat ccatgaagcg gacgtggtcg tcgaagaaat aacccaggtc tggtttgtcg
      721 gatttgccca taccgatcag gtctggagca atgcagcgat gggtcggtgc aacatgcggg
      781 atgatgttgc gccacacgta ggaggaggtc gggttaccgt gcaggaacag cacaggggtg
      841 ccatcgcgcg gaccaacatc gacgtagtgc atgcgctcgc ccaggacttc cacataatgg
      901 gggtcgaatg gaaagccagt accgatttct gccatatggc taccacgagg caccaggccg
      961 ctgctgtgat gatgatgatg atggctgctg cccatggtat atctccttct taaagttaaa
     1021 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
     1081 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
     1141 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
     1201 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
     1261 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
     1321 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
     1381 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     1441 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     1501 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     1561 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     1621 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     1681 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     1741 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     1801 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     1861 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     1921 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     1981 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     2041 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     2101 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     2161 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     2221 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     2281 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     2341 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     2401 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     2461 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2521 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     2581 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     2641 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     2701 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     2761 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     2821 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     2881 gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct
     2941 ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg
     3001 actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg
     3061 tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca
     3121 tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc
     3181 attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
     3241 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
     3301 ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag
     3361 tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa
     3421 cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc
     3481 ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg
     3541 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     3601 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     3661 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     3721 gattgtactg agagtgcacc attgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     3781 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3841 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3901 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3961 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4021 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4081 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4141 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4201 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4261 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4321 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4381 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4441 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4501 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4561 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4621 cgttaaggga ttttggtcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     4681 gggtgttatg agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa
     4741 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     4801 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     4861 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     4921 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     4981 cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     5041 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     5101 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     5161 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     5221 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     5281 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     5341 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     5401 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     5461 gaataaattg cagtttcatt tgatgctcga tgagtttttc taagaattaa ttcatgagcg
     5521 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5581 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5641 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5701 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5761 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5821 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     5881 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5941 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     6001 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     6061 aatccggata tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca
     6121 aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct
     6181 ttgttactag tttttcttaa catctggctt cttacttttg ggaacaagga aaacattgcc
     6241 atctcttgtt tgctgcaggg agtattcact aggagaataa ggttttccat cttcatcacg
     6301 tagcatgctg aaaacttcga gatataaggt gctgagttgt tttttcagta ggtgaaggct
     6361 tttgtcattt tctccttttt ctttgagcaa tttttctttt tcatctttca aatgatctaa
     6421 atcttgctct agttctacta tattttccag ttttcttttt ctgcaattct gagcagccac
     6481 tttattctta cccctcctac gtatatcccg aattaatgca agttgagctt cattgaactg
     6541 ctctttggac atcatttcgt tgaagtcaac aacagggagg ttaatgattt tttctacagg
     6601 gaatgggata tggagagctt ttgccctaag ttcatctctt gtgagatgag cctccaagcg
     6661 gcttgaatgt ttgtcttttg tgaatggggt tttccgatga ccaggactta caggcaattc
     6721 tttctctggt gtgttctcac attgggcatc atgcacgtga gtgctctgcc cctgagatgg
     6781 tgacaagggt tgtaccatat ccccagaaga atgtactggt gttttaggac cattctgttt
     6841 gacacttcca ggggcactat ctagctcttc cacttcagaa tcactgaggc caagtagtgt
     6901 gtctccatag ctggaagatt ccactgagtg ttctggtgat gccacactgg gacttgtgtt
     6961 tagtgaaatg ccggagtcag aatcattgaa ttctgctgtg ctttcagggt ggttttggtt
     7021 gaaagctttg caaagtgata gatcagaaac atcaatgggc ccatttagaa gttcagagag
     7081 tgaatggctt aaagtagcag gtgagggcat gctgttgctg atactgggct cagctatgaa
     7141 agcagaataa aattcatcac caaaatctgt gttgactgtg gcatctgaat ttaatgagtt
     7201 cactgtcaac tggttggggt cttctgtgga gaggatgctg ctgaaggaat cctcaaaagc
     7261 attaagaaaa tgtggactac agttacctac ttctttttcc attgagggta tagatgagta
     7321 aaaatgataa ttgtcaactt ctgtcagttt ggcttctgga cttggaacca tggtagtctc
     7381 aaccagcttg tcattttcaa tattaagaca ctgtaactca ggaatggata atagctcctc
     7441 ccaaacttgc tcaatgtcct gttgcatacc gtctaaatca acaggggcta cctgagcaac
     7501 agaagtttca ggtgactgag cctgattagt agcaatgaag actgggctct cgatgtgacc
     7561 gggaatatca ggaacaagtg actgaaacgt agccgaagaa acctcattgt catctacaaa
     7621 cgggaatgtc tgcgccaaaa gctgcatgca gtcatcaaag tacaaagcat ctgatttggg
     7681 aatgtgggca acctgggagt agttggcaga tccactggtt tctgactgga tgtgctgggc
     7741 tggctgaatt gggagaaatt cacctgtctc ttcatctagt tgtaactgag cgaaaaaggc
     7801 tttctcttgc tccttttgga gttgttcttg tctttccttt tcaagttttt tctgtttttc
     7861 cagctcatac tctttccgtc gctgactgaa gtcaaatact tctcgactta ctccaagatc
     7921 tatatcttgc ctccaaagta tgtcaatcaa atccatgtcc tgctgggacg ggagtcccgg
     7981 cggcggcagc tcc
//
